Allosteric properties of phosphate-activated glutaminase of human liver mitochondria.

The kinetics of human liver phosphate-activated glutaminase were studied in mitochondria isolated from surgical biopsies. The pH profile and activation by phosphate closely resembled rat liver glutaminase and differed clearly from human or rat kidney mitochondrial glutaminases. The activity responses to glutamine or phosphate were allosteric, showing positive cooperativity, as in the rat liver enzyme. Exogenous 1 mM NH4Cl shifted the glutamine concentration at half-maximal velocity, [Gln]0.5, to lower values without changing Vmax or sigmoidicity. Hill plots showed a parallel shift to the left with NH4Cl and the apparent number of binding sites, nH, was 2-3. 25 mM KHCO3 gave the same effects as NH4Cl on [Gln]0.5, Vmax, sigmoidicity and nH. The combination of the two activators was less than additive. Glutamate did not inhibit. We postulate that liver glutaminase is allosteric in its kinetics because it plays a key role in urea synthesis by regulating provision of glutamate for synthesis of N-acetylglutamate, the obligatory co-factor of carbamoylphosphate synthetase.